2011 ADA Posters 1261-2041.indd - Diabetes

saline-treated group fed ad lib. CTL ASO did not change BW or BF versus
saline-CR. However, R4 ASO further lowered BW by 7-11% and BF by 23-
25%. In addition, R4 ASO reduced epididymal fat pad wt by 16-22% and
peri-renal fat pad wt by 29-33% without effect on lean mass. As expected,
CR signifi cantly reduced whole body VO 2 . In contrast, R4 ASO prevented the
decline in VO 2 . In separate studies, R4 ASO Rx of lean C57BL/J and CD-1
mice for 3 mo resulted in > 75% reduction in liver FGFR4 mRNA and was well
tolerated without any overt side effects, indicating lack of mechanism based
toxicity with chronic reduction of FGFR4. To further explore mechanism of
action, the effect of R4 ASO Rx on FGF15 levels was determined. Four-wk
Rx of DIO mice reduced liver FGFR4 mRNA by > 75%, increased ileum FGF15
mRNA by > 5-fold and plasma FGF15 protein levels by > 2-fold. These rodent
fi ndings were confi rmed in Cynomolgus monkeys, where Rx with R4 ASO for 3
mo reduced liver FGFR4 mRNA by 70% and caused > 5-fold increase of ileum
FGF19 mRNA and 2-fold increase of plasma FGF19 protein levels without any
overt toxicities. These data provide further support that peripheral inhibition
of FGFR4 with antisense drugs is an attractive therapeutic approach for
obesity.
1823-P
Caloric Restriction Reverses Elevated ER Stress and Oxidative
Stress in Liver of ob/ob Mice
ATSUYUKI TSUTSUMI, HIROYUKI MOTOSHIMA, SHUJI KAWASAKI, SATOKO
HANATANI, MOTOYUKI IGATA, TATSUYA KONDO, TAKESHI MATSUMURA, KAKU
TSURUZOE, TAKESHI NISHIKAWA, EIICHI ARAKI, Kumamoto, Japan
Endoplasmic reticulum (ER) stress and oxidative stress have been
proposedto play a crucial role in the development of insulin resistance
and diabetes in obesity. Although caloric restriction (CR) improves various
obesity-related disorders, the effects of CR on ER stress and oxidative stress
in obesity have not been elucidated.
To investigate how CR affects ER stress, oxidative stress and insulin
signaling in obesity, a leptin-defi cient ob/ob mice under CR (ob-CR) or ad
libitum (AL)–feeding (ob-AL) for four weeks were compared on daily foodintake,
body weight (BW), glucose metabolism, ER stress, oxidative stress
and insulin signaling. To reduce BW to the level of lean littermates fed AL
(lean-AL), the ob-CR were given reduced chow (2.0 g/day) to a level which
previously reported to extend lifespan of ob/ob mice. Glucose and insulin
tolerance, markers for ER stress and oxidative stress (protein nitrotyrosine),
and insulin signaling were investigated in these animals.
CR reduced BW in ob-CR, resulted in comparable BWs between ob-CR
and lean-AL after 2 weeks, both of which became smaller than ob-AL in BW.
The ob-CR showed improved glucose tolerance and hepatic insulin action
compared with ob-AL. Signifi cant increases in ER stress markers (such as
phosphorylation of PERK and eIF2α, and expression of ATF4 and GRP78
mRNA) and in protein nitrotyrosine were observed in liver and epididymal
fat from ob-AL compared with those from lean-AL. CR signifi cantly reduced
all of these markers in ob/ob mice. CR also signifi cantly reduced both serinephosphorylation
of IRS-1 and JNK phosphorylation in liver of ob/ob mice.
Reduction in ER stress by CR tended to be stronger than that obtained by
treatment with 4-phenyl butyric acid, which is known to reduce ER stress
in vivo.
In conclusion, four weeks of CR effectively reduced ER stress and oxidative
stress, and improved insulin action via suppression of JNK-mediated IRS-1
phosphorylation in liver of ob/ob mice.
1824-P
Catechol-o-Methyltransferase Defi ciency Is Associated with Ab normal
Adiposity and Glucose Intolerance: Implication for a New Mechanism
of Metabolic Syndrome
MEGUMI KANASAKI, KEIZO KANASAKI, DAISUKE KOYA, Uchinada, Japan
Catechol-o-methyltransferase (COMT), an enzyme linked with several
neuronal/psychological diseases, metabolizes a group of catechols including
catecholamines and catechol estrogens. 2-methoxyestradiol, one of the
metabolite of catechol estrogens via COMT, exerts as an anti-angiogenesis
and a vascular protective factor via inhibition of hypoxia-inducible factor
(HIF)-1α. Several single-nucleotide polymorphisms (SNPs) in the human
COMT gene have been observed and some of which may result in the
depletion of catalytic activity, such as the well-known functional Val158Met
isoform. These COMT SNPs have been associated with hypertension,
preeclampsia, obesity, increased fat mass and obesity in diabetes. However,
the contribution of COMT suppression in the onset of metabolic diseases
has not been determined. Using COMT-inhibitor (Ro 41-0960), here we
show that COMT-inhibitor accelerate adiposity with the impaired glucose
tolerance in mice fed a high-fat diet. A 2-week high-fat diet (60% fat in total
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energy) induced impaired glucose tolerance when analyzed by the intraperitoneal
glucose tolerance test in mice. COMT-inhibitor treatment for last
1 week displayed the exacerbation of glucose tolerance defects and insulin
resistance in mice with a high-fat diet. Such impaired glucose tolerance
was associated with the hepatosteatosis, decreased liver glycogen level
associated with HIF-1α and macrophage accumulation in epididymal fat. A
signifi cant angiogenesis in the mesenteric fat was also observed in highfat
fed mice treated with COMT-inhibitor. These abnormal adiposity and
glucose tolerance defect with the abnormal angiogenesis in COMT-inhibitor
treated mice were ameliorated by the 2-ME treatment. 2-ME suppressed
HIF-1α and macrophage accumulation in the epididymal fat. These results
indicated that 2-ME could be the potential target drug for the treatment of
metabolic defects via inhibition of abnormal adipogenesis, angiogenesis and
infl ammation especially in low COMT genotype population.
Supported by: Kanae Foundation (K.K.)
1825-P
Cathepsin K, L Expression in Adipocytes Is Upregulated by Palmitate
Via TNFα and IL-6
JEONG SEON YOO, YOUNGMI LEE, EUN HAE LEE, JIWOON KIM, SHINYOUNG
LEE, JANG-HAN JUNG, JI SUN NAM, SHIN AE KANG, TAE-WOONG NOH, MIN
HO CHO, JONG SUK PARK, KYUNG WOOK KIM, JAE-WOO KIM, CHUL WOO AHN,
BONG SOO CHA, EUN JIG LEE, SUNG KIL LIM, KYUNG RAE KIM, HYUN CHUL LEE,
Seoul, Republic of Korea
Aim: Cathepsin family is lysosomal cysteine protease. It is recently
reported that cathepsin K, L, S control adipogenesis and relate to acute
coronary syndrome. However, it is not clear whether cathepsins expression
can be affected by saturated fatty acid which has a critical role in metabolic
disease. We examined the hypothesis that palmitate upregulates cathepsin
K, L, S expression in obese adipose tissue and infi ltrating macrophages have
crucial role in this process via infl ammatory cytokines.
Methods and Results: 3T3-L1 cells were fully differentiated to mature
adipocytes for 8 to 10 days and treated with palmitate, lipopolysaccharide
(LPS) which is ligand of TLR4 (toll-like receptor 4). Real-time PCR revealed that
not cathepsin S but cathepsin K, L expression is upregulated by palmitate as
dose- and time-dependent manner. But there was no additional effect when
we use palmitate-treated RAW264.7 cells’ media instead of direct treatment
to 3T3-L1 cells. Cathepsin K, L is upregulated after treatment of TNFa or IL-6
like palmitate. Six-week-old C3H/HeN mice with normal TLR4 and C3H/HeJ
mice with TLR4 gene’s mutation got LPS injection ip (1 mg/kg) or were fed
with high fat diet (60% fat of total calories) for 13 weeks and sacrifi ce to
harvest epididymal fat. Cathepsin K, L, S expression increased in both kinds
of mice fed high fat diet compared with chow diet. But we cannot fi nd similar
pattern at mice which got LPS ip injection.
Conclusions: We concluded that cathepsin K, L expression in fat tissue are
upregulated by saturated fatty acid via infl ammatory cytokines, not TLR4,
thereby might have a critical role in developing acute coronary syndrome
of metabolic syndrome and there are no additional effects of infi ltrating
macrophages.
1826-P
Comparison of Genetic Factors Controlling Abdominal Fat Distribution
and Glucose Metabolism in A/J and SM/J Mice
MISATO KOBAYASHI, TAMIO OHNO, MASAKO KUGA, MASAHIKO NISHIMURA,
ATSUSHI MURAI, FUMIHIKO HORIO, Nagoya, Japan
Obesity is a major risk factor for insulin resistance, type 2 diabetes,
dys lipidemia, cardiovascular disease, fatty liver and stroke. However,
each abdominal fat depot, such as mesenteric or epididymal, differently
contributes to the development of insulin resistance. Previously, we
mapped a major quantitative trait locus (QTL, T2dm2sa) for impaired glucose
tolerance on chromosome (Chr.) 2 and revealed that the chromosomal region
near T2dm2sa on Chr. 2 had affective genes on not only glucose tolerance,
but also the accumulation of body fat, using SM.A-T2dm2sa congenic
mice. SM.A-T2dm2sa mice possess the A/J-allele T2dm2sa region in SM/J
mice. To identify the genetic regions that contribute to fat accumulation in
epididymal and mesenteric fat depots and to examine whether or not the
genetic regions that affect glucose metabolism and body fat distribution
are coincident, we performed QTL analyses using (A/J×SM.A-T2dm2sa)F2
intercross mice. SM.A- T2dm2sa mice shows lower glucose tolerance and
higher epididymal fat weight compared with A/J mice.
Signifi cant QTLs for mesenteric fat were detected on Chrs. 5 (Mfatq3sa,
LOD: 3.5), 7 (Mfatq1sa, LOD: 7.7) and 16 (Mfatq2sa, LOD: 5.0). Signifi cant
QTLs for epididymal fat were detected on Chrs. 10 (Efatq2sa, LOD: 3.8) and
For author disclosure information, see page 785.
Integrated Physiology/
Obesity
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